Purification of phospholamban, a 22,000 dalton protein from cardiac sarcoplasmic reticulum that is specifically phosphorylated by cyclic AMP-dependent protein kinase
Very low concentrations deoxycholate (DOC) were used to isolate two proteins from canine cardiac sarcoplasmic reticulum. These two proteins are phospholamban, a 22,000 dalton protein, and the Ca/sup 2 +/ + Mg/sup 2 +/-ATPase, the major protein of the sarcoplasmic reticulum, responsible for the active transport of calcium. The 22,000 dalton protein is first solubilized in a very low concentration of DOC and then subjected to column chromatography. After molecular weight sieving on a Sephadex G-75 column, the 22,000 dalton protein appears as a purified protein on sodium dodecyl sulfate (SDS)-polyacrylamide gels. The purified protein is specifically phosphorylated by cyclic AMP-dependent protein kinase. Phospholipids are still bound to the isolated protein. The Ca/sup 2 +/ + Mg/sup 2 +/-ATPase is purified by first solubilizing all the extrinsic proteins with a low concentration of DOC. An increasing amount of DOC is then added to yield the purified Ca/sup 2 +/ + Mg/sup 2 +/-ATPase. This protein is at least 95% pure. Adding additional DOC to the purified enzyme enhances the enzyme's ability to hydrolyze ATP. (ERB)
- Research Organization:
- Rochester Univ., NY (USA). Dept. of Radiation Biology and Biophysics
- DOE Contract Number:
- AC02-76EV03490
- OSTI ID:
- 5008458
- Report Number(s):
- UR-3490-1805
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ATP-ASE
PURIFICATION
PROTEINS
PHOSPHORYLATION
DETERGENTS
DOGS
ELECTROPHORESIS
ENZYME ACTIVITY
EXPERIMENTAL DATA
HEART
IN VITRO
LIQUID COLUMN CHROMATOGRAPHY
MUSCLES
PHOSPHOLIPIDS
PHOSPHOTRANSFERASES
ADDITIVES
ANIMALS
BODY
CARDIOVASCULAR SYSTEM
CHEMICAL REACTIONS
CHROMATOGRAPHY
DATA
EMULSIFIERS
ENZYMES
ESTERASES
ESTERS
HYDROLASES
INFORMATION
LIPIDS
MAMMALS
NUMERICAL DATA
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHOSPHATASES
SEPARATION PROCESSES
SURFACTANTS
TRANSFERASES
VERTEBRATES
WETTING AGENTS
550200* - Biochemistry